Qflow cytometry-based receptoromic screening: A high-throughput quantification approach informing biomarker selection and nanosensor development

Si Chen, Jared Weddell, Pavan Gupta, Grace Conard, James Parkin, Princess I. Imoukhuede

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Nanosensor-based detection of biomarkers can improve medical diagnosis; however, a critical factor in nanosensor development is deciding which biomarker to target, as most diseases present several biomarkers. Biomarker-targeting decisions can be informed via an understanding of biomarker expression. Currently, immunohistochemistry (IHC) is the accepted standard for profiling biomarker expression. While IHC provides a relative mapping of biomarker expression, it does not provide cell-by-cell readouts of biomarker expression or absolute biomarker quantification. Flow cytometry overcomes both these IHC challenges by offering biomarker expression on a cell-by-cell basis, and when combined with calibration standards, providing quantitation of biomarker concentrations: this is known as qFlow cytometry. Here, we outline the key components for applying qFlow cytometry to detect biomarkers within the angiogenic vascular endothelial growth factor receptor family. The key aspects of the qFlow cytometry methodology include: antibody specificity testing, immunofluorescent cell labeling, saturation analysis, fluorescent microsphere calibration, and quantitative analysis of both ensemble and cell-by-cell data. Together, these methods enable high-throughput quantification of biomarker expression.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages117-138
Number of pages22
DOIs
StatePublished - Jan 1 2017

Publication series

NameMethods in Molecular Biology
Volume1570
ISSN (Print)1064-3745

Keywords

  • Angiogenesis
  • Background subtraction
  • Heterogeneity
  • Immuno-labeling
  • Mixture modeling
  • Platelet-Derived Growth Factor (PDGF)
  • QFlow cytometry
  • Quantitative flow cytometry
  • Systems biology
  • Vascular Endothelial Growth Factor (VEGF)

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Chen, S., Weddell, J., Gupta, P., Conard, G., Parkin, J., & Imoukhuede, P. I. (2017). Qflow cytometry-based receptoromic screening: A high-throughput quantification approach informing biomarker selection and nanosensor development. In Methods in Molecular Biology (pp. 117-138). (Methods in Molecular Biology; Vol. 1570). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6840-4_8